论文部分内容阅读
目的:研究羟基喜树碱是否可通过线粒体途径诱导人肝癌细胞SMMC-7721凋亡.方法:将不同浓度的羟基喜树碱作用于肝癌细胞SMMC-7721,用MTT法检测细胞增殖,光镜观察细胞形态;用荧光染料MitocaptureTM检测线粒体跨膜电位、用Western blot法检测细胞色素C从线粒体的释放;用AnnexinⅤ-FITC,hoechst法及电镜检测细胞的凋亡状况.结果:羟基喜树碱对肝癌细胞SMMC-7721的增殖有明显的抑制作用,IC50约是80 mg/L.当用80 mg/L羟基喜树碱作用不同时间后:线粒体膜跨电位下降并伴随有细胞色素C从线粒体至细胞质的释放;细胞膜的磷脂酰丝氨酸外翻;细胞核染色质固缩,呈致密浓染的凋亡状态.结论:羟基喜树碱通过线粒体途径引起线粒体跨膜电位下降与细胞色素C的释放可能是其诱导肿瘤细胞凋亡的途径之一.
Aims: To study whether hydroxycamptothecin can induce apoptosis in SMMC-7721 cells through mitochondria pathway.Methods: HCPT cells were treated with different concentrations of HCPT in SMMC-7721 cells and the cell proliferation was detected by MTT assay. Cell morphology was detected by fluorescent dye MitocaptureTM.The cytochrome C release from mitochondria was detected by Western blot.Apoptosis was detected by AnnexinⅤ-FITC, hoechst and electron microscopy.Results: The proliferation of SMMC-7721 cells was significantly inhibited with an IC50 of about 80 mg / L. After 80 mg / L hydroxycamptothecin was used for different time periods, the mitochondrial membrane decreased transmutationally and accompanied cytochrome C from mitochondria to cytoplasm Release of cytosolic phosphatidylserine, pyknosis of nuclear chromatin, and dense apoptosis of apoptotic cells.Conclusion: The decrease of mitochondrial transmembrane potential and the release of cytochrome C by hydroxycamptothecin may be the result of mitochondrial pathway One of the ways to induce tumor cell apoptosis.